Identification of genes regulating stimulus-dependent synaptic assembly in Drosophila using an automated synapse quantification system

• Published in: Genes & Genetic Systems Vol. 97; no. 6; pp. 22-00114 - 309
• Main Authors: Osaka, Jiro, Yasuda, Haruka, Watanuki, Yusuke, Kato, Yuya, Nitta, Yohei, Sugie, Atsushi, Sato, Makoto, Suzuki, Takashi
• Format: Journal Article
• Language: English
• Published: Japan The Genetics Society of Japan 01-12-2022; Japan Science and Technology Agency
• Subjects: active zone; Animals; Automation; Axons; Axons - metabolism; Drosophila; Drosophila - genetics; Drosophila melanogaster - genetics; Drosophila Proteins - genetics; Drosophila Proteins - metabolism; Drosophila visual system; Feasibility studies; Image processing; Insects; Latrophilin; Neural plasticity; Neurons - metabolism; Photoreceptors; Protein interaction; Receptors, Peptide - metabolism; RNA-mediated interference; Synapses - metabolism; Synaptic plasticity; synaptic remodeling; Latrophilin; Drosophila visual system; synaptic remodeling; active zone
• ISSN: 1341-7568; 1880-5779
• DOI: 10.1266/ggs.22-00114

Neural activity-dependent synaptic plasticity is an important physiological phenomenon underlying environmental adaptation, memory and learning. However, its molecular basis, especially in presynaptic neurons, is not well understood. Previous studies have shown that the number of presynaptic active zones in the Drosophila melanogaster photoreceptor R8 is reversibly changed in an activity-dependent manner. During reversible synaptic changes, both synaptic disassembly and assembly processes were observed. Although we have established a paradigm for screening molecules involved in synaptic stability and several genes have been identified, genes involved in stimulus-dependent synaptic assembly are still elusive. Therefore, the aim of this study was to identify genes regulating stimulus-dependent synaptic assembly in Drosophila using an automated synapse quantification system. To this end, we performed RNAi screening against 300 memory-defective, synapse-related or transmembrane molecules in photoreceptor R8 neurons. Candidate genes were narrowed down to 27 genes in the first screen using presynaptic protein aggregation as a sign of synaptic disassembly. In the second screen, we directly quantified the decreasing synapse number using a GFP-tagged presynaptic protein marker. We utilized custom-made image analysis software, which automatically locates synapses and counts their number along individual R8 axons, and identified cirl as a candidate gene responsible for synaptic assembly. Finally, we present a new model of stimulus-dependent synaptic assembly through the interaction of cirl and its possible ligand, ten-a. This study demonstrates the feasibility of using the automated synapse quantification system to explore activity-dependent synaptic plasticity in Drosophila R8 photoreceptors in order to identify molecules involved in stimulus-dependent synaptic assembly.